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鸡集落刺激因子(CSF)基因的克隆、序列分析及蛋白结构预测

DOI: 10.7668/hbnxb.2009.02.008, PP. 36-41

Keywords: 克隆,,集落刺激因子,序列分析,蛋白质结构预测

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Abstract:

根据GenBank中报道的红原锦鸡集落刺激因子(Colony-stimulatingfactor,CSF)cDNA序列,利用PrimerPremier5.0软件设计一对特异性引物,对本地肉鸡注射100μg/mLConA试剂,饲养24h后,取脾脏提取淋巴细胞总RNA,利用RT-PCR技术克隆集落刺激因子(CSF)的cDNA片段.对克隆片段进行测序,并利用DNAstar软件进行不同物种间同源性比较.序列分析表明,CSF的cDNA开放阅读框为435bp,编码144个氨基酸,与红原锦鸡的序列比较,其核苷酸的同源性为98.4%,氨基酸的同源性为95.2%,同人、犬、印度野牛、野猪、绵羊和老鼠的CSF序列作比较,其核苷酸的同源性分别为28.7%,29.2%,33.%,30.8%,29.4%和34.7%,氨基酸的同源性分别为8.3%,9.0%,15.3%,12.4%,11.0%和12.0%.然后利用DNAstar和DNAman软件,对鸡CSF的结构进行预测,结果表明,鸡CSF基因为一结构松散的蛋白分子.鸡的CSF基因被成功克隆,它存在着种属特异性.这为进一步研究该基因的生物学作用特别是利用CSF增强DNA疫苗的免疫效果奠定了基础.

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